本文關(guān)鍵詞：膠囊滲透壓泵控釋IGFBPrP1對(duì)小鼠肝肺組織的影響　出處：《山西醫(yī)科大學(xué)》2011年碩士論文　論文類(lèi)型：學(xué)位論文

  更多相關(guān)文章： 纖維化 肝臟 肺臟 胰島素樣生長(zhǎng)因子結(jié)合蛋白相關(guān)蛋白1 膠囊滲透壓泵

【摘要】：研究背景： 胰島素樣生長(zhǎng)因子結(jié)合蛋白相關(guān)蛋白1 (insulin-like growth-factor binding protein-related protein 1, IGFBPrP1)是一種可溶性細(xì)胞黏附糖蛋白,由腫瘤細(xì)胞、血管內(nèi)皮細(xì)胞、平滑肌細(xì)胞和成纖維細(xì)胞等多種細(xì)胞分泌,Lopez-Bermejo等和Ma等的研究發(fā)現(xiàn)IGFBPrP1蛋白廣泛表達(dá)于人體各種組織細(xì)胞并起著相當(dāng)重要的作用,能夠調(diào)節(jié)上皮細(xì)胞和纖維母細(xì)胞生長(zhǎng)、增殖、黏附,促進(jìn)血管形成,刺激內(nèi)皮細(xì)胞產(chǎn)生前列環(huán)素,誘導(dǎo)細(xì)胞凋亡等多種功能。目前有關(guān)IGFBPrP1蛋白功能尚不是完全清楚。導(dǎo)師劉立新課題研究小組發(fā)現(xiàn)：用重組IGFBPrP1能使體外培養(yǎng)的肝星狀細(xì)胞株HSC-T6活化,且在一定的劑量范圍內(nèi)隨著IGFBPrP1劑量的增加HSC的活化程度逐漸增強(qiáng)；抗IGFBPrP1抗體通過(guò)抑制肝纖維化小鼠肝臟HSC的活化、減少ECM的生成及減少肝細(xì)胞的凋亡等途徑發(fā)揮抗纖維化作用。IGFBPrP1可促進(jìn)體外培養(yǎng)的肝星狀細(xì)胞過(guò)表達(dá)TGF-β1及細(xì)胞外基質(zhì),增多的細(xì)胞外基質(zhì)是由TGFβ1通過(guò)Smad3信號(hào)通路介導(dǎo)的。IGFBPrP1可能是肝纖維化發(fā)生發(fā)展過(guò)程中的種新的因子。 IGFBPrP1的生物學(xué)功能還有待進(jìn)一步研究：一方面,腹腔注射動(dòng)物給藥IGFBPrP1半衰期短,有藥物濃度高峰,代謝快,不能真正了解IGFBPrP1蛋白的功能；另一方面,IGFBPrP1有致肝纖維化的作用,對(duì)其他臟器是否也有影響及其可能機(jī)制是什么也尚未明確。為了闡明此問(wèn)題,設(shè)計(jì)了本課題。 目的： 比較IGFBPrP1對(duì)小鼠肝、肺組織作用的異同,探討IGFBPrP1對(duì)小鼠肝、肺組織的影響及其可能機(jī)制。 方法： 24只3周齡SPF級(jí)雄性C57 BL/6野生型小鼠,體重(12±2)g,由中國(guó)醫(yī)學(xué)科學(xué)院實(shí)驗(yàn)動(dòng)物研究所提供。隨機(jī)分為3組：手術(shù)組(8只)用1%水合氯醛麻備小鼠,于小鼠后背部皮下行一0.5厘米左右橫切口,組織鉗由切口處皮下向小鼠后背部撐出一個(gè)膠囊大小的空間(空間應(yīng)大到膠囊泵體可以有一定移動(dòng)度,但不能讓泵體輕易滑到小鼠側(cè)翼),行手術(shù)植入注滿100μg/ml IGFBPrP1溶液的膠囊滲透壓泵(輸藥管釋出口要朝內(nèi)塞入皮下不可反向以致釋出口剛好在傷口之下)。膠囊泵連續(xù)恒定釋放IGFBPrP1溶液,泵容積90μ1,泵速0.11μ1/h,持續(xù)4周。假手術(shù)組(8只)行單純對(duì)照手術(shù)切開(kāi)縫合不植入膠囊泵,正常組(8只)不作處理。在第4周末禁食10h后,乙醚麻醉采集肝、肺組織用10%中性甲醛固定,石蠟包埋,切片。(1)分別對(duì)肝、肺組織石蠟切片做HE染色和苦味酸-天狼猩紅染色觀察肝、肺組織病理變化及膠原纖維形成情況；(2)免疫組織化學(xué)染色檢測(cè)肝、肺組織中IGFBPrP1、III型膠原(Collagen III)、Smad3及p-Smad2/3的表達(dá)；(3) TUNEL法檢測(cè)各組肝組織中肝細(xì)胞凋亡情況。 結(jié)果： (1)苦味酸-天狼猩紅染色結(jié)果：手術(shù)組肝組織中膠原纖維含量較正常組和假手術(shù)組明顯增多(0.46%±0.09%vs1.27.%±0.10%,0.45%±0.08%vs1.27.%±0.10%,F=597.797,P0.01),手術(shù)組肺組織中膠原纖維含量較其他兩組差異不明顯(0.35%±0.09%vs0.36%±0.12%,0.37%±0.12%vs 0.36%±0.12%,F=0.344,P0.05)。 (2)免疫組織化學(xué)染色結(jié)果：手術(shù)組肝組織中IGFBPrP1、III型膠原(CollagenIII)及p-Smad2/3的表達(dá)均較正常組和假手術(shù)組顯著增強(qiáng),Smad3表達(dá)未見(jiàn)增強(qiáng)。(IGFBPrP1: 0.26±0.04 vs 1.77±0.64,0.25±0.04 vs 1.77±0.64, F=44.064, P0.01; Collagen III:0.45±0.04 vs 2.59±0.29,0.43±0.05 vs 2.59±0.29, F=431.641,P0.01; p-Smad2/3:0.53±0.07 vs 2.44±0.32,0.51±0.08vs2.44±0.32, F=114.251, P0.01; Smad3:0.29±0.06 vs 0.33±0.04, 0.34±0.04 vs 0.33±0.04, F=1.928, P0.05)。手術(shù)組肺組織中IGFBPrP1 III型膠原(Collagen III)、Smad3及P-Smad2/3的表達(dá)較其他兩組差異不明顯(IGFBPrP1:0.32±0.05vs0.30±0.07, 0.29±0.06vs0.30±0.07, F=0.505, P0.05; Collagen III:0.93±0.07 vs 0.89±0.02,0.85±0.01 vs0.89±0.02, F=0.973, P0.05; Smad3:0.26±0.03 vs 0.25±0.04,0.24±0.09 vs 0.25±0.04, F=0.150, P0.05; p-Smad2/3:0.25±0.06 vs 0.26±0.05,0.28±0.06 vs 0.26±0.05, F=0.255, P0.05)。 (3) TUNEL法檢測(cè)肝細(xì)胞凋亡結(jié)果：熒光顯微鏡下和普通光鏡下手術(shù)組肝組織中凋亡肝細(xì)胞較正常組和假手術(shù)組明顯增多(6.21%土2.83%vs 34.14%±11.04%,7.13%±2.59%vs34.14%士11.04%,F=114.251,P0.01；6.10%士2.23%vs 32.23%±10.60%,5.71%±2.87%vs32.23%±10.60%,F=114.251,P0.01)。 結(jié)論： (1)持續(xù)恒量的外源性IGFBPrP1可致小鼠肝纖維化,同等條件下對(duì)小鼠肺組織影響甚微而不致肺纖維化。 (2)IGFBPrP1對(duì)小鼠組織臟器的作用具有選擇性。
[Abstract]:Research background:
Insulin-like growth factor binding protein related protein 1 (insulin-like growth-factor binding protein-related protein 1, IGFBPrP1) is a soluble cell adhesion glycoprotein by tumor cells, vascular endothelial cells, smooth muscle cells and fibroblasts into a variety of cell secretion, Lopez-Bermejo and Ma, and plays an important role in widespread expression the IGFBPrP1 protein in various human tissues and cells, can regulate the growth of epithelial cells and fibroblast proliferation, adhesion, angiogenesis, endothelial cells before the column ring element functions, inducing cell apoptosis. The IGFBPrP1 protein function is not entirely clear. Professor Liu Lixin research group found that recombinant IGFBPrP1 can make in vitro hepatic stellate cells activation of HSC-T6, and in a certain dose range with increasing dose of IGFBPrP1 The activation level of HSC increased gradually; the anti IGFBPrP1 antibody by inhibiting the activation of hepatic fibrosis in HSC mice, reduced the production of ECM and reduce the apoptosis of liver cells exerts anti fibrosis effect of.IGFBPrP1 can promote hepatic stellate cells in vitro expression of TGF- beta 1 and extracellular matrix, the extracellular matrix is increased by TGF beta 1 via Smad3 signal pathway mediated by.IGFBPrP1 may be in the process of the development of a new factor of hepatic fibrosis.
The biological function of IGFBPrP1 remains to be further studied: on the one hand, intraperitoneal injection of animal drug IGFBPrP1 has short half-life, peak drug concentration, fast metabolism and the function of IGFBPrP1 protein can not really understand; on the other hand, IGFBPrP1 induced hepatic fibrosis, to other organs have the effect and the possible mechanism is also not yet clear what in order to clarify this issue, the design of this topic.
Objective:
The effects of IGFBPrP1 on the liver and lung tissue of mice were compared, and the effect of IGFBPrP1 on the liver and lung tissue of mice and its possible mechanism were discussed.
Method錛，

本文編號(hào)：1408844